Limits...
CLE19 expressed in the embryo regulates both cotyledon establishment and endosperm development in Arabidopsis.

Xu TT, Ren SC, Song XF, Liu CM - J. Exp. Bot. (2015)

Bottom Line: CLE19 is expressed in the epidermal layers of the cotyledon primordia, hypocotyl, and root cap in the embryo.Transgenic plants carrying an antagonistic CLE19 G6T construct expressed under the control of CLE19 regulatory elements exhibited a dominant seed abortion phenotype, with defective cotyledon establishment in embryos and delayed nuclear proliferation and cellularization in endosperms.We therefore propose that CLE19 may act as a mobile peptide co-ordinating embryo and endosperm development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Fragrant Hill, Beijing 100093, China University of Chinese Academy of Sciences, Beijing 100049, China.

No MeSH data available.


Related in: MedlinePlus

Expression of cotyledon-specific genes in arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants. (A–D) Confocal microscopic examinations of embryos excised from wild-type plants (WT) carrying pWOX1:SV40-3XGFP (A) or pWOX3:SV40-3XGFP marker constructs (C), or from plants carrying pWOX1:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 (B) or pWOX3:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 double constructs (D). Note that similar GFP expression, though with a reduced level, was observed in cotyledon primordia of arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T), as compared with embryos from plants carrying only marker constructs (A, C). Scale bars=50 μm. (E, F) qRT-PCR showed reduced levels of WOX1 (E) and WOX3 expression (F) in ovules from the wild type (WT) and pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T). Data represent the mean ±SD from three independently extracted RNA samples. Asterisks indicate significant differences from the wild type (P < 0.01 by Student’s t-test).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4526921&req=5

Figure 5: Expression of cotyledon-specific genes in arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants. (A–D) Confocal microscopic examinations of embryos excised from wild-type plants (WT) carrying pWOX1:SV40-3XGFP (A) or pWOX3:SV40-3XGFP marker constructs (C), or from plants carrying pWOX1:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 (B) or pWOX3:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 double constructs (D). Note that similar GFP expression, though with a reduced level, was observed in cotyledon primordia of arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T), as compared with embryos from plants carrying only marker constructs (A, C). Scale bars=50 μm. (E, F) qRT-PCR showed reduced levels of WOX1 (E) and WOX3 expression (F) in ovules from the wild type (WT) and pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T). Data represent the mean ±SD from three independently extracted RNA samples. Asterisks indicate significant differences from the wild type (P < 0.01 by Student’s t-test).

Mentions: To elucidate further whether the defect in cotyledon development in pCLE19:CLE19G6T:tCLE19 plants is attributed to failures in cotyledon initiation or cotyledon establishment, two cotyledon marker lines were developed based on published in situ hybridization results for WUSCHEL-RELATED HOMEOBOX 1 (WOX1) and WOX3 (Haecker et al., 2004). Upstream sequences from WOX1 and WOX3 (4454bp and 5028bp, respectively) were fused to the SV40-3XGFP reporter gene and transformed into the wild-type Arabidopsis (Col-0). Transgenic plants were examined and crossed to pCLE19:CLE19G6T:tCLE19 plants, and progeny plants carrying pCLE19:CLE19G6T:tCLE19 and homozygous pWOX1:SV40-3XGFP or pWOX3:SV40-3XGFP constructs were examined under CLSM. As shown in Fig. 5, GFP expression in torpedo-stage embryos from wild-type plants carrying pWOX1:SV40-3XGFP was restricted to cells located at the edges of two cotyledons (Fig. 5A). Embryos from plants carrying both the pCLE19:CLE19G6T:tCLE19 and the pWOX1:SV40-3XGFP constructs exhibited a similar GFP expression pattern despite a severe delay in cotyledon development in these embryos (Fig. 5B). GFP expression in wild-type torpedo-stage embryos carrying the pWOX3:SV40-3XGFP construct was observed at the adaxial side of two cotyledons (Fig. 5C). Similar GFP expression was also observed in transgenic plants carrying both the pWOX3:SV40-3XGFP and pCLE19:CLE19G6T:tCLE19 constructs (Fig. 5D). Real-time PCR analyses confirmed that both WOX1 and WOX3 were expressed in pCLE19:CLE19G6T:tCLE19 plants, albeit with a 30–40% reduction observed in ovules carrying defective embryos compared with those from the wild type (Fig. 5E, F). These data suggest that cotyledon initiation was unaffected in pCLE19:CLE19G6T:tCLE19 transgenic plants.


CLE19 expressed in the embryo regulates both cotyledon establishment and endosperm development in Arabidopsis.

Xu TT, Ren SC, Song XF, Liu CM - J. Exp. Bot. (2015)

Expression of cotyledon-specific genes in arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants. (A–D) Confocal microscopic examinations of embryos excised from wild-type plants (WT) carrying pWOX1:SV40-3XGFP (A) or pWOX3:SV40-3XGFP marker constructs (C), or from plants carrying pWOX1:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 (B) or pWOX3:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 double constructs (D). Note that similar GFP expression, though with a reduced level, was observed in cotyledon primordia of arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T), as compared with embryos from plants carrying only marker constructs (A, C). Scale bars=50 μm. (E, F) qRT-PCR showed reduced levels of WOX1 (E) and WOX3 expression (F) in ovules from the wild type (WT) and pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T). Data represent the mean ±SD from three independently extracted RNA samples. Asterisks indicate significant differences from the wild type (P < 0.01 by Student’s t-test).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4526921&req=5

Figure 5: Expression of cotyledon-specific genes in arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants. (A–D) Confocal microscopic examinations of embryos excised from wild-type plants (WT) carrying pWOX1:SV40-3XGFP (A) or pWOX3:SV40-3XGFP marker constructs (C), or from plants carrying pWOX1:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 (B) or pWOX3:SV40-3XGFP/pCLE19:CLE19G6T:tCLE19 double constructs (D). Note that similar GFP expression, though with a reduced level, was observed in cotyledon primordia of arrested embryos from pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T), as compared with embryos from plants carrying only marker constructs (A, C). Scale bars=50 μm. (E, F) qRT-PCR showed reduced levels of WOX1 (E) and WOX3 expression (F) in ovules from the wild type (WT) and pCLE19:CLE19G6T:tCLE19 transgenic plants (CLE19G6T). Data represent the mean ±SD from three independently extracted RNA samples. Asterisks indicate significant differences from the wild type (P < 0.01 by Student’s t-test).
Mentions: To elucidate further whether the defect in cotyledon development in pCLE19:CLE19G6T:tCLE19 plants is attributed to failures in cotyledon initiation or cotyledon establishment, two cotyledon marker lines were developed based on published in situ hybridization results for WUSCHEL-RELATED HOMEOBOX 1 (WOX1) and WOX3 (Haecker et al., 2004). Upstream sequences from WOX1 and WOX3 (4454bp and 5028bp, respectively) were fused to the SV40-3XGFP reporter gene and transformed into the wild-type Arabidopsis (Col-0). Transgenic plants were examined and crossed to pCLE19:CLE19G6T:tCLE19 plants, and progeny plants carrying pCLE19:CLE19G6T:tCLE19 and homozygous pWOX1:SV40-3XGFP or pWOX3:SV40-3XGFP constructs were examined under CLSM. As shown in Fig. 5, GFP expression in torpedo-stage embryos from wild-type plants carrying pWOX1:SV40-3XGFP was restricted to cells located at the edges of two cotyledons (Fig. 5A). Embryos from plants carrying both the pCLE19:CLE19G6T:tCLE19 and the pWOX1:SV40-3XGFP constructs exhibited a similar GFP expression pattern despite a severe delay in cotyledon development in these embryos (Fig. 5B). GFP expression in wild-type torpedo-stage embryos carrying the pWOX3:SV40-3XGFP construct was observed at the adaxial side of two cotyledons (Fig. 5C). Similar GFP expression was also observed in transgenic plants carrying both the pWOX3:SV40-3XGFP and pCLE19:CLE19G6T:tCLE19 constructs (Fig. 5D). Real-time PCR analyses confirmed that both WOX1 and WOX3 were expressed in pCLE19:CLE19G6T:tCLE19 plants, albeit with a 30–40% reduction observed in ovules carrying defective embryos compared with those from the wild type (Fig. 5E, F). These data suggest that cotyledon initiation was unaffected in pCLE19:CLE19G6T:tCLE19 transgenic plants.

Bottom Line: CLE19 is expressed in the epidermal layers of the cotyledon primordia, hypocotyl, and root cap in the embryo.Transgenic plants carrying an antagonistic CLE19 G6T construct expressed under the control of CLE19 regulatory elements exhibited a dominant seed abortion phenotype, with defective cotyledon establishment in embryos and delayed nuclear proliferation and cellularization in endosperms.We therefore propose that CLE19 may act as a mobile peptide co-ordinating embryo and endosperm development.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Fragrant Hill, Beijing 100093, China University of Chinese Academy of Sciences, Beijing 100049, China.

No MeSH data available.


Related in: MedlinePlus